Chromatographic-mass Spectrometric Technique Research Articles

Unveiling the photostability of the molecule of colour shikonin: From solution to the solid state

Shikonin, Shk, is a molecule of colour characterized by its natural phenolic-based compound composition, belonging to the naphthoquinone family. It is known for its low solubility in water and limited resistance to lightfastness. Shk is present in variable amount in the dried roots of Lithospermum erythrorhizon, Lithospermum officinale, and Alkanna tinctoria. The dyestuff extracted from these plants has historically been employed in Asia and Europe for textile dyeing, producing various shades of purple. Despite the historical significance of this dye, it has received limited attention in the context of cultural heritage studies. In the present work, Shk and model hydroxynaphthoquinone dyes were irradiated with polychromatic and monochromatic light in solution and in the solid state, mimicking paint mock-ups, to evaluate their photochemical stability. Low photoreaction quantum yield values (ϕR < 10−5) were observed in solution, providing quantitative evidence for the high stability of this molecule to light exposure. Chromatographic-mass spectrometric analyses carried out on the irradiated solutions and extracts from artificially aged Shk paint mock-ups, detected 5,8-dihydroxy-2-(1‑hydroxy-3-oxo-4-methyl-4-pentenyl)-l,4-naphthoquinone as degradation product, together with newly identified intermediate species involved in the complex photodegradation process. These indicate that, both in solution and in the solid state, the photoreaction of Shk proceeds with a similar mechanism. In the context of heritage science, this work contributes to highlight the potentialities of the adopted noninvasive approach, corroborated by results collected by chromatographic-mass spectrometric techniques, in identifying Shk in historical and archaeological specimens through its absorption and emission features and/or by detecting its degradation products which are, for the first time, characterized in terms of their UV–Vis absorption and emission spectral properties.

Development and validation of the chromatography-mass spectrometry technique for polyprenols quantitative assessment

Introduction. Polyprenols are known as a class of natural long-chain isoprenoid alcohols, which are natural bioregulators that directly participate in the synthesis of cell membrane glycoproteins. Their hepatoprotective activity is proven, as well other types of their pharmacological effects are known, which is the reason of significant interest in these substances as a promising medicinal product. It is non-trivial task to determine the sum of polyprenols in extracts as include design and implementation of accurate reproducible analytical methods, which will subsequently be used in standardization.Aim. Development and validation of the chromatographic-mass spectrometric technique for polyprenols identification and their quantitative assessment.Materials and methods. Chromatographic separation of polyprenols was performed by using an HPLC Agilent 1260 Infinity II (Agilent Technologies, США); with the mixture of methanol, n-hexane, propanol-2, and aqueous ammonium acetate solution as eluent in gradient mode. An AB Sciex QTrap® 3200MD (AB Sciex Pte. Ltd., Singapore) triple quadrupole mass spectrometer was used as a detector, with the registration of polyprenols adducts.Results and discussion. The conditions for chromatographic separation and detection of polyprenols were identified. The developed technique was validated for the following characteristics: specificity, limit of detection, limit of quantification, linearity, accuracy, precision, range of application, and stability.Conclusion. It was determined the content of polyprenols in the substance recieved from Ginkgo biloba L. and Picea abies L. The developed technique can be used in the future to assess the content of polyprenols in drug products or pharmaceutical substances.

AN LC-ESI-MS/MS METHOD DEVELOPMENT AND VALIDATION FOR THE QUANTIFICATION OF INFIGRATINIB IN BIOLOGICAL MATRICES

Objective: The study was aimed to develop a precise and simple liquid chromatographic electrospray ionization tandem mass spectrometric (LC-ESI-MSMS) technique is essential for the quantification of Infigratinib in biological matrices. Methods: Chromatographic resolution was attained with PhenominexC18 (50 mm×2.6 mm, 3 µm) stationary column and a mobile solvent composition of 0.1% HCOOH, methyl alcohol and acetonitrile in the proportion of 10:10:80. Chromatograms were resolved by an isocratic separation with a flowing rate of 0.50 ml/min at 40 °C. Results: Quantitation was executed by monitoring the transitions of m/z. 560.19/189.13 for Infigratinib and 494.5→394.5 for Imatinib internal standard in multiple reaction monitoring. The standard curve regression line was y = 0.0016x+0.0062 and the correction coefficient (r2) was 0.9994. The % CV outcomes for matrix effect at Lower-QC and Higher-QC were 4.95% and 3.61% respectively. The percentage average recoveries for Infigratinib in Higher-QC (900ng/ml), MQC (600ng/ml) and Lower-QC (3ng/ml) were 93.27%, 94.69% and 97.24% respectively. The intra and interday precisions of analytical procedure was estimated by assessing the %CV outcomes and were in between 1.88 to 5.93% for the QC samples. Conclusion: The developed procedure can be useful for the assessment of Infigratinib in biological matrices in quality control, forensic and bioavailability studies.

Stability Indicating an LC-MS/MS Method Development and Validation for the Quantification of Capmatinib in Human Plasma

A linear and accurate liquid chromatographic tandem mass spectrometric (LC-MS/MS) technique for the quantitation of capmatinib in plasma was developed and subjected for validation. Canagliflogin was employed as an internal standard (IS). Extraction of plasma was executed utilizing 5 mL of ethyl acetate solvent. Analysis was performed Zorbax ODS (50 mm × 2.1 mm x 3 μ) stationary phase at room conditions and a movable solvent composition of 0.1% HCOOH, acetonitrile and methyl alcohol (15:50:35). The flowing rate of the movable phase was 0.4 mL/min. Drug and IS were identified in positive mode of ionization with electrospray mode to get the mass transitions of (m/z): Capmatinib, 413.15/386.14 and canagliflogin (IS), 445.15/267.12. The correlation between capmatinib concentrations and their respective peak proportions to canagliflogin was a straight line over 0.2 to 3200 ng/mL concentration levels. Intra day and inter-day precisions were ≤5.18% for capmatinib. Inter and intra day bias were within the range of −4.17 to 4.53%. The mean measured extraction recovery of capmatinib was 99.23%. Recovery of IS was 98.31%. Capmatinib was subjected for long-term, freeze thaw, bench top, short-term stability, auto-sampler, dry extract, and stock solution stability at low QC and high QC levels and it was stable at all these conditions. The established technique can be utilized to regularly quantitfy of capmatinib in plasma samples in industries, forensic labs, and clinical research organizations.

Response Surface Methodology in Optimising the Extraction of Polyphenolic Antioxidants from Flower Buds of Magnolia × soulangeana Soul.-Bod. var. 'Lennei' and Their Detailed Qualitative and Quantitative Profiling.

A response surface methodology (RSM) with a central composite design (CCD) was developed to predict and apply the best ultrasound-assisted extraction (UAE) conditions, including the extraction time, the composition of aqueous-ethanolic extractants, and the solvent-to-plant-material ratio, for obtaining the highest yields of different types of polyphenolic components from the dried flower buds of Magnolia × soulangeana Soul.-Bod. var. 'Lennei' (MSL). The novel approach in the RSM procedure resulted from the simultaneous optimisation of UAE conditions to obtain extracts with the highest antioxidant and antiradical potential (examined as dependent variables), using appropriate spectrophotometric assays, with Folin-Ciocâlteu and 2,2-diphenyl-1-picrylhydrazyl reagents, respectively. The use of 66.8% (V/V) ethanol as the extraction solvent during the 55.2 min extraction protocol and the ratio of extractant volume to herbal substance of 46.8 mL/g gave the highest total yield of bioactive antioxidant phenolics in the extract obtained. For this herbal preparation, a qualitative and quantitative analysis was performed using combined chromatographic (LC), spectroscopic (PDA), and tandem mass spectrometric (ESI-QToF-MS/MS) techniques. A detailed phytochemical profiling, conducted for the first time, documented substantial amounts of various polyphenolic antioxidants, especially phenylethanoids and flavonoids, in the MSL flower buds. Their average total content exceeded 30.3 and 36.5 mg/g dry weight, respectively.

Volatilomics of raspberry fruit germplasm by combining chromatographic and direct-injection mass spectrometric techniques.

The application of direct-injection mass spectrometric (DI-MS) techniques, like Proton Transfer Reaction Time of Flight Mass Spectrometry (PTR-ToF-MS) has been suggested as a reliable phenotyping tool for fruit volatilome assessment in both genetic and quality-related studies. In this study the complexity of raspberry aroma was investigated by a comprehensive untargeted VOC analysis, done by combining SPME-GC-MS and PTR-ToF-MS assessments with multi-block discriminant analysis using the DIABLO mixOmics framework. The aim was to acquire an exhaustive characterization of the raspberry volatilome according to different fruit ripening stages (pink, ripe, and overripe) and genetic variances (50 accessions), as well as to investigate the potential of PTR-ToF-MS as a rapid and high throughput VOC phenotyping tool to address issues related to raspberry fruit quality. Results of this study demonstrated the complementarity between SPME-GC-MS and PTR-ToF-MS techniques to evaluate the raspberry aroma composition. PTR-ToF-MS generates reliable raspberry VOC fingerprints mainly due to a reduced compound fragmentation and precise content estimation. In addition, the high collinearity between isomers of monoterpenes and norisoprenoids, discovered by GC analysis, reduces the main analytic limitation of PTR-ToF-MS of not being able to separate isomeric molecules. The high similarity between the VOC matrices obtained by applying PTR-ToF-MS and SPME-GC-MS confirmed the possibility of using PTR-ToF-MS as a reliable high throughput phenotyping tool for raspberry volatiolome assessment. In addition, results provided by the germplasm collection investigation enabled to distinguish the best performing accessions, based on VOCs composition, to be used as superior parental lines for future breeding programs.

Non-target estrogenic screening of 60 pesticides, six plant protection products, and tomato, grape, and wine samples by planar chromatography combined with the planar yeast estrogen screen bioassay

For non-target residue analysis of xenoestrogens in food, sophisticated chromatographic–mass spectrometric techniques lack in biological effect detection. Various in vitro assays providing sum values encounter problems when opposing signals are present in a complex sample. Due to physicochemical signal reduction, cytotoxic or antagonistic effect responses, the resulting sum value is falsified. Instead, the demonstrated non-target estrogenic screening with an integrated planar chromatographic separation differentiated opposing signals, detected and prioritized important estrogenic compounds, and directly assigned tentatively the responsible compounds. Sixty pesticides were investigated, ten of which showed estrogenic effects. Exemplarily, half-maximal effective concentrations and 17β-estradiol equivalents were determined. Estrogenic pesticide responses were confirmed in six tested plant protection products. In food, such as tomato, grape, and wine, several compounds with an estrogenic effect were detected. It showed that rinsing with water was not sufficient to remove selected residues and illustrated that, though not usually performed for tomatoes, peeling would be more appropriate. Though not in the focus, reaction or breakdown products that are estrogenic were detected, underlining the great potential of non-target planar chromatographic bioassay screening for food safety and food control.Graphical

Wastewater-based epidemiology combined with forensic toxicological information: trends in drug use and impact of law enforcement activities

Wastewater-based epidemiology (WBE) has recently become one of the most rapidly evolving disciplines engaged in the measurement of population-level illicit drug use. Still, limited evidence has been presented how population-level WBE data correlates to forensic toxicological statistics and law enforcement activities. Objective was to study (1) how WBE methodology can efficiently give early-warning type of information on drug use and its trends, (2) how WBE findings compared to number of drivers under influence of drugs (DUID), and (3) how (exceptionally large) law enforcement operation affected actual drug use. Wastewater analysis: 24-h wastewater influent composite samples ( n = 190) were collected during 04/2013–03/2022 from Helsinki wastewater treatment plant (WWTP) serving approximately 860,000 inhabitants. Furthermore, several national sampling campaigns up to 28 WWTPs covering approximately 60% of all Finnish population (3.2 million out of 5.5 million) during 2012–2022 were performed. All samples were analysed by validated UHPLC-MS/MS. For comparison, traffic cases: Prevalence of different drugs was studied in all suspected DUID cases in Finland (from 2013 to 2021). All cases (whole blood) were systematically analysed by chromatographic mass spectrometric techniques. Law enforcement operation of illicit drugs: Information on Operation Greenlight (Trojan Shield), which was an exceptional operation in size together with Finnish and several international law enforcement authorities worldwide starting in February 2021 in Finland. Both wastewater analysis data and suspected DUID cases confirm the significant rise of amphetamine and cocaine use between 2013 to early 2021 in Finland. Based on WBE, amphetamine use peaked at record-high level from mid-2020 (after COVID-19 lockdown) to spring 2021 in the capital area of Finland. Mean amphetamine mass loads were 92 mg/1000 persons/day in 2013, 335 in 2020, 322 in H1/2021, 234 in H2/2021 and 162 in Q1/2022 at the wastewater of Helsinki WWTP. The use of amphetamine significantly decreased in 2021 after the start of Operation Greenlight, while for cocaine effects on population-level use were more temporary. The number of amphetamine positive DUID in Finland showed similar trend: 2064 cases in 2013 (45.9% of all suspected DUID cases), 7913 (65.8%) in 2020 and 6228 (63.6%) in 2021 for amphetamine. For cocaine, the similar figures in wastewater were 18 (2013), 119 (2020), 110 (H1/2021), 144 (H2/2021) and 141 mg/1000 persons/day (Q1/2022), respectively. The number of cocaine-related DUID increased from 81 (2013) to 501 (2021). Cocaine use is still highly concentrated in the capital area of Finland and there has been 6-8-fold increase in cocaine use at population-level during the last eight years. Based on WBE, the use of amphetamine was partially replaced by methamphetamine and synthetic cathinones, such as alfa-PHP, at the end of 2021, but not close to the same extent as amphetamine use was reduced. The results show in detail how use of various illicit drugs has changed during the last decade in Finland. WBE and DUID results have strongly correlated with each other concerning drug use and its trends: increase/decrease in population-level drug use has also increased/decreased harmful societal phenomena of illicit drugs, such as DUID. The results obtained during the last year indicate that unexceptionally large and successful law enforcement operations may affect drug supply, reduce actual drug use and diminish harmful societal effects, such as DUID. Extensive 10-year data and experience on WBE approach shows that objective, novel and early-warning type of information on drug use and its trends for various illicit drugs can be obtained. More comprehensive picture and unique perspectives on drug use and drug markets are further obtained when combining different indicators and information, as exemplified here with WBE and DUID data linked to law enforcement activities.

Production and identification of intricate bioactive oligosaccharides from Nyctanthesarbor-tristis leaves by a combination of enzymatic, HPAEC and MALDI-TOF-MS techniques

Nondigestible oligosaccharides are associated with positive health outcomes; however, hemicellulosic oligosaccharides of N. arbor-tristis have not been studied. Herein we report production hemicellulosic oligosaccharides from its leaves and their identification. A series of complex neutral oligosaccharides (1-16), produced by endo-(1,4)-b-d-cellulase digestion of the alkali extracted xyloglucan, were identified by chemical, HPAE chromatographic and MALDI mass spectrometric techniques. Additionally, xylan enriched hemicellulose-A fraction upon endo-(1,4)-b-d-xylanase digestion yielded several acidic and neutral xylo-oligosaccharides (17-24), each containing a backbone of b-(1,4)-d-Xylp units substituted at position 2 with a single stub of d-Xylp and 4-O-Me-d-GlcpA residues. The structural diversity and high-water solubility of these enzyme-generated oligosaccharides implied promising health implication.

Evaluation of chemical profiles and biological properties of Gliricidia sepium (Jacq.) Walp. volatile oils from Nigeria

Volatile oils obtained by hydrodistillation from Gliricidia sepium (Jacq.) Walp. leaf and stem were examined for their chemical composition and biological activity. The oils were analyzed by gas chromatographic-mass spectrometric (GC-MS) techniques. Toxicity, antimicrobial and antioxidant activities were evaluated via brine shrimp lethality assay, agar-diffusion, and DPPH radical-scavenging methods, respectively. A total of 43 and 44 constituents were identified in the leaf and stem oils, correspondingly. The major components recognized in the leaf oil were (E)-hexadecatrienal (16.9%) and pentadecanal (16.0%) while humulene epoxide II (17.5%) and caryophyllene oxide (10.6%) dominated the stem oil. There was no significant activity against the bacteria but moderate inhibition zones (IZ) between 1.8±0.3 and 8.1±0.1 mm were observed against the fungi. The stem oil showed better antifungal activity than the leaf oil but not as active as the standard drug ketoconazole which inhibited the test fungi with IZ range of 10.4±0.4-21.0±1.4 mm at 200 μg. Both oils were toxic to brine shrimp (Artemia salina) giving LC50 of 79.7 μg/mL (leaf) and 38.7 μg/mL (stem). The leaf and stem oils had IC50 of 84.3 μg/mL and 142.2μg/mL, respectively, in the DPPH radical-scavenging assay, indicating moderate antioxidant activity relative to positive controls, butylated hydroxylanisole (IC50=45.1 μg/mL) and α-tocopherol (IC50 =81.6 μg/mL). The results suggest that G. sepium volatile oil may find potential use as a natural antioxidant and antifungal agent.

Application of an innovative metabolomics approach to discriminate geographical origin and processing of black pepper by untargeted UHPLC-Q-Orbitrap-HRMS analysis and mid-level data fusion

An untargeted metabolomics approach based on ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS) fingerprinting was applied to investigate the metabolic differences of black pepper among three geographical origins (Sri Lanka, Vietnam, and Brazil) and two post-harvest processing (sterilized and non-sterilized spice). Principal component analysis (PCA) was employed to assess the overall clustering of samples, whereas supervised orthogonal partial least squares discriminant analysis (OPLS-DA) was effectively used for discrimination purposes. OPLS-DA models were fully validated (R2Y and Q2 values > 0.5) and the variable importance in projection (VIP) approach was employed to provide valuable data about differential metabolites with high discrimination potential (8 markers were putatively identified). For origin differentiation, three markers were highlighted with VIP values > 1.5 (i.e. reynosin, artabsinolide D, and tatridin B). Fatty acid derivates were the most frequent markers within the metabolites annotated for processing discrimination (e.g. 10,16-dihydroxyhexadecanoic acid and 9-hydroperoxy-10E-octadecenoic acid). Additionally, different combinations of mid-level data fusion of chromatographic-mass spectrometric techniques (UHPLC and gas chromatography coupled to HRMS) and proton nuclear magnetic resonance spectroscopy (1H NMR) were evaluated for the first time for geographical and processing discrimination of black pepper. The NMR-UHPLC-GC mid-level fused model was preferred among the tested fusion approaches since good sample clustering and no misclassification were achieved. Enhanced correct classification rate was achieved by mid-level data fusion compared with the findings obtained for one of the individual techniques (1H NMR fingerprinting) (from 92% to 100% of samples correctly classified). This study opens the path to new metabolomics approaches for black pepper authentication and quality control.

Synthetic Cannabinomimetics: A Brief History and the Challenges They Pose for the Forensic Chemist

Since the first detection of synthetic cannabinomimetics in herbal smoking blends in 2008 the clandestine production of these compounds, based on seizure data, increased in number every year until ~2012. In recent years there has been a decline in synthetic cannabinomimetic production both in number and diversity. The synthetic details of the first generation cannabinoids were documented in the scientific and medical literature making production comparatively simple. Subsequent generations of synthetic cannabinoids involved more complex but still very practicable synthetic chemistry. This resulted in a period of rapid growth in synthetic cannabinoids creating a health crisis and problems for forensic chemists faced with many substances for which no certified reference materials existed. Routine forensic chemistry laboratories were well practiced at identifying known drugs using chromatographic–mass spectrometric techniques and comparison to reference materials. However as synthetic cannabinomimetics, often referred to in the literature as synthetic cannabinoids, appeared in large numbers, few laboratories were equipped with the nuclear magnetic resonance (NMR) spectrometers and high resolution mass spectrometers (HRMS) required for identification of unknown substances. These developments also challenged public prosecutors for opinions from forensic experts as to the legality or otherwise of these novel drugs.

Extensive Chemometric Investigations of Distinctive Patterns and Levels of Biogenic Amines in Fermented Foods: Human Health Implications.

Although biogenic amines (BAs) present in fermented foods exert important health-promoting and physiological function support roles, their excessive ingestion can give rise to deleterious toxicological effects. Therefore, here we have screened the BA contents and supporting food quality indices of a series of fermented food products using a multianalyte-chemometrics strategy. A liquid chromatographic triple quadrupole mass spectrometric (LC-MS/MS) technique was utilized for the simultaneous multicomponent analysis of 8 different BAs, and titratable acidity, pH, total lipid content, and thiobarbituric acid-reactive substances (TBARS) values were also determined. Rigorous univariate and multivariate (MV) chemometric data analysis strategies were employed to evaluate results acquired. Almost all foods analyzed had individual and total BA contents that were within recommended limits. The chemometrics methods applied were useful for recognizing characteristic patterns of BA analytes and food quality measures between some fermented food classes, and for assessing their inter-relationships and potential metabolic sources. MV analysis of constant sum-normalized BA profile data demonstrated characteristic signatures for cheese (cadaverine only), fermented cod liver oil (2-phenylethylamine, tyramine, and tryptamine), and wine/vinegar products (putrescine, spermidine, and spermine). In conclusion, this LC-MS/MS-linked chemometrics approach was valuable for (1) contrasting and distinguishing BA catabolite signatures between differing fermented foods, and (2) exploring and evaluating the health benefits and/or possible adverse public health risks of such products.

Er:YAG laser cleaning of a marble Roman urn

There are very few documented instances of the use of an Er:YAG laser in the cleaning of stone artefacts; however, the cases reported in the literature have shown its effectiveness in the removal of surface organic patinas and lichens. A Roman marble funerary urn (67–100 AD) of archaeological importance was covered with an intractable surface layer that obscured its delicate decorative carving. Chromatographic mass spectrometric and Raman spectroscopic techniques allowed the characterisation of those inorganic and organic materials that had undergone photo-oxidation and biological changes, resulting in the thick encrustation. An Er:YAG laser at 2940nm, with a pulse length of 300µsec, was used for cleaning the urn. Preliminary tests were performed with fluences ranging from 0.7 to 6.4J/cm2 and in the presence of a variety of wetting agents. The laser cleaning resulted in a sufficiently clean marble surface with the thick deposit of unwanted material removed. Finally, the sensitivity of the method allowed for a compact layer of calcium oxalate to be retained on the marble’s surface to help protect it.

Assessment of Metabolites Content of Native Alcoholic Beverages Obtained from Jos, Nigeria

Burukutu, (BKT) Pito (PT) and Goskolo (GSK) are drinks consumed routinely within the area of study. Although consumption of GSK is illegal owing to its life threatening toxicity, people still consume it secretly. Whereas BKT and PT are locally prepared using unscientific methods in preparing the drinks, GSK is a mixture containing different compounds and chemicals. BKT and PT are more commonly ingested than GSK. Because methods applied during ‘brewing’ BKT and PT are not scientific, toxic metabolites might be generated. So, the drinkers are prone to their toxic effects. This work ascertained the presence and levels of unwanted metabolites in these drinks using Gas chromatographic-Mass spectrometric (GC-MS) technique. Metabolites could aggravate the toxicity of alcohol and alcohol-related disease conditions. Results, (% relative abundance), showed the significant presence in BKTof6-Octadecanoic acid (38.51), Hexadecanoic acid (20.92), Isopentyl alcohol (12.27), 1,11,13-Octadecatriene (11.60) among others. In PT,9-Octadecanoic acid (39.40), Hexadecanoic acid (15.98), Carbonic acid methylpentyl ester (14.22), 9,17-Octadecadienal (6.37) were the major components. In GSK, 6-Octadecenoic acid (42.43), n-Hexadecanoic (17.51), 1-Butanol (16.52) 9 and 9,12-Octadecadienoic acid (10.72) were the dominant components. BKT, PT, and GSK are composed of compounds/metabolites that are organic in nature, with about 83% of them being organic acids. This leads to a decrease in pH thereby making the drinks acidic. It is concluded that the alcoholic beverages contain metabolites that predispose drinkers to metabolic or respiratory acidosis, due to drop in pH. Furthermore, these metabolites could aggravate the toxicity of alcohol. Other components of the samples include ketones, different types of alcohols, aldehydes, derivatives of benzene and esters.

Metabolite Profiling by Hyphenated Liquid Chromatographic Mass Spectrometric Technique (HPLC-DAD-ESI-Q-TOF-MS/MS) and Neurobiological Potential of Haplophyllum sahinii and H. vulcanicum Extracts.

In the current study, the ethanol extracts of flower, stem, and root parts of two endemic Turkish species, e. g., Haplophyllum sahinii O. Tugay & D. Ulukuş and H. vulcanicum Boiss. & Heldr., were screened against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) associated with Alzheimer's disease as well as tyrosinase (TYR) linked to Parkinson's disease using ELISA microplate assay at 200 μg/mL. Among the extracts, the highest inhibition was caused by the stem extract of H. sahinii against BChE (IC50 =64.93±1.38 μg/mL). Consistently, all of the extracts were found to exert a selective inhibition towards BChE to some extent. It was only the root extract of H. vulcanicum that could inhibit AChE at low level (IC50 =203.18±5.33 μg/mL). None of the extracts displayed an inhibition over 50 % against TYR. Metabolite profiling of the extracts was achieved by a highly hyphenated liquid chromatographic mass spectrometric technique (HPLC-DAD-ESI-Q-TOF-MS/MS), which revealed the presence of furoquinoline (β-fagarine, γ-fagarine) and amide (tubasenicine, tubacetine) alkaloids; furano- (rutamarin), pyrano- (xanthyletine), and geranyloxy coumarins; phenylpropanoid (secoisolariciresinol), arylnaphthalene (mono-O-acetyldiphyllin apioside), and dibenzylbutyrolactone (kusunokinin, haplomyrfolin) lignans. Several important differences were observed between the extracts analyzed. β-Fagarine was the major alkaloid in H. vulcanicum, whereas γ-fagarine was present only in the roots of both Haplophyllum species; moreover, secoisolariciresinol and secoisolariciresinol dimethyl ether were the main lignans in the stems and flowers. This is the first study identifying ChE and TYR inhibitory effect and metabolic profiles of H. vulcanicum and H. sahinii.

Gas Chromatographic Study of Bio-active Compounds in Methanolic Extract of Leaf of Crateva adansonii DC

This study investigates the volatile bioactive components in Crateva adansonii plant that has been applied in the treatment of diverse ailments in the Western part of Nigeria and in Africa. The methanolic leaf extract of Crateva adansonii was subjected to analysis using gas chromatographic-mass spectrometric technique. The mass spectra of the compounds were analysed and identity of the compounds confirmed using the data base of the National Institute of Standard and Technology (NIST) library. The compounds identified includesn-Hexadecanoic acid; 9,12-Octadecadienoic acid (Z,Z); 9-Octadecenoic acid; (Z,E)-2,13-Octadecadien-1-ol; 9,17-Octadecadienal; cis-9-Hexadecenal; cis-Vaccenic acid; Z,Z-10,12-Hexadecadienal; 13-Octadecenal; Hexadecanoic acid 2-hydroxy-1-(hydroxymethyl)ethyl ester; E-11-Hexadecenal. Most of the compounds identified have antibacterial, anti-oxidant, anti-inflammatory and hypocholesterolemic properties, which affirms the therapeutic applications of Crateva adansonii as a medicinal plant.

Photo-oxidation processes of Rhodamine B: A chromatographic and mass spectrometric approach

The study and characterization of organic dyes in textiles, wastewaters, food, cosmetics and forensic specimens is an increasingly important field of research and a challenge for analytical chemistry. Sensitive and specific methods are required to determine molecular markers at very low concentrations and to detect and identify their relative degradation products. In this field, the present paper deals with the development of an extremely sensitive analytical strategy able to identify synthetic organic dyes even after ageing, such as photo-oxidation resulting in severe fading, by identifying specific stable markers. Rhodamine B (C.I. 45,170), belonging to the class of xanthenes, was chosen as representative of fugitive dyes of high commercial and historical importance, in the frame of a research promoted by the European platform IPERION-CH. The behaviour of Rhodamine B used as a dyestuff for wool yarns, subjected to accelerated photo-ageing, was investigated by chromatographic, spectroscopic and high resolution mass spectrometric techniques. The ultra-sensitive analytical method developed, based on core shell reverse phase analytical column separation, allowed us to characterize Rhodamine B oxidation products investigating their mass fragmentation and rationalizing the resulting ageing pathway. These degradation compounds, characterized in artificially aged reference wool yarns, confirmed to be valuable markers for the identification of Rhodamine B even in a complex historical textile matrix as a red Florentine velvet cope subjected to restoration in the 20th century.

Exploring Blueberry Aroma Complexity by Chromatographic and Direct-Injection Spectrometric Techniques.

Blueberry (Vaccinium spp.) fruit consumption has increased over the last 5 years, becoming the second most important soft fruit species after strawberry. Despite the possible economic and sensory impact, the blueberry volatile organic compound (VOC) composition has been poorly investigated. Thus, the great impact of the aroma on fruit marketability stimulates the need to step forward in the understanding of this quality trait. Beside the strong effect of ripening, blueberry aroma profile also varies due to the broad genetic differences among Vaccinium species that have been differently introgressed in modern commercial cultivars through breeding activity. In the present study, divided into two different activities, the complexity of blueberry aroma was explored by an exhaustive untargeted VOC analysis, performed by two complementary methods: SPME-GC-MS (solid phase microextraction- gas chromatography-mass spectrometry) and PTR-ToF-MS (proton transfer reaction-time of flight-mass spectrometry). The first experiment was aimed at determining the VOC modifications during blueberry ripening for five commercially representative cultivars (“Biloxi,” “Brigitta Blue,” “Centurion,” “Chandler,” and “Ozark Blue”) harvested at four ripening stages (green, pink, ripe, and over-ripe) to outline VOCs dynamic during fruit development. The objective of the second experiment was to confirm the analytical capability of PTR-ToF-MS to profile blueberry genotypes and to identify the most characterizing VOCs. In this case, 11 accessions belonging to different Vaccinium species were employed: V. corymbosum L. (“Brigitta,” “Chandler,” “Liberty,” and “Ozark Blue”), V. virgatum Aiton (“Centurion,” “Powder Blue,” and “Sky Blue”), V. myrtillus L. (three wild genotypes of different mountain locations), and one accession of V. cylindraceum Smith. This comprehensive characterization of blueberry aroma allowed the identification of a wide pull of VOCs, for the most aldehydes, alcohols, terpenoids, and esters that can be used as putative biomarkers to rapidly evaluate the blueberry aroma variations related to ripening and/or senescence as well as to genetic background differences. Moreover, the obtained results demonstrated the complementarity between chromatographic and direct-injection mass spectrometric techniques to study the blueberry aroma.

Multidimensional Liquid Chromatography Coupled with Tandem Mass Spectrometry for Identification of Bioactive Fatty Acyl Derivatives.

Recognition of the contributions of lipids to cellular physiology, both as structural components of the membrane and as modulatory ligands for membrane proteins, has increased in recent years with the development of the biophysical and biochemical tools to examine these effects. Their modulatory roles in ion channels and transporters function have been extensively characterized, with the molecular mechanisms of these activities being the subject of intense scrutiny. The physiological significance of lipids in biochemistry is expanding as numerous fatty acyls are discovered to possess signaling properties. These bioactive lipids are often found in quantities of pmol/g of tissue and are co-extracted with numerous lipophilic molecules, making their detection and identification challenging. Common analytical methodologies involve chromatographic separation and mass spectrometric techniques; however, a single chromatographic step is typically ineffective due to the complexity of the biological samples. It is, therefore, essential to develop approaches that incorporate multiple dimensions of separation. Described in this manuscript are normal phase and reversed phase separation strategies for lipids that include detection of the bioactive primary fatty acid amides and N-acyl glycines via tandem mass spectrometry. Concerted utilization of these approaches are then used to separate and sensitively identify primary fatty acid amides extracted from homogenized tissue, using mouse brains as a test case.